Mitigating Antibiotic Resistance

UFlorida iGEM 2021

TABLE OF

CONTENTS

Introduction

What is iGEM? Who are we?

Our Project Part 1

Our goals, background information and relevant

context

Our Project Part 2

The Science Stuff!

How you can get involved!

You can contribute to our project!

01 Introduction

What is iGem? Who are we?

iGEM stands for Internationally Genetically

Engineered Machine

What is iGEM?

PRIZES!

Best Foundational Advance Project

Best Environment Project

Best Energy Project

Best Information Processing Project

Best Hardware Project

Best Food & Nutrition Project

Best Diagnostics Project

Grand prizes!

Best Manufacturing Project

So… what is an iGEM team?

There are multiple components!

WET LAB

DRY LAB

HUMAN PRACTICES

WIKI

WET LAB

DRY LAB

HUMAN PRACTICES

WIKI

WHO ARE WE?

Scientists

Programmers

Engineers

College of Liberal Arts and Sciences

College of Engineering

Artists

02 Our Project Part 1

Background

Information

Our Goals

We wanted to tackle a problem...

So we just had several Zoom meetings where we brainstormed!

ANTIBIOTIC RESISTANCE!

So what is that exactly?

Antibiotic Resistance is...

Antibiotic Resistance threatens everyone...

Vocabulary alert!

Antibiotic Resistance

all

Antimicrobial Resistance

mean

Drug Resistance

the same thing!

WHERE DOES IT COME FROM?

Quick review on evolution...

1. Individuals in a population show variation

2. Variations can be inherited

3. Organisms have more offspring that can survive under normal circumstances

4. Variations that increase reproductive success will have a greater chance of being passed

on from generation to generation

5. A population will slowly change over time in response to the environment

So in terms of evolution...

● Individuals in a population show variation → not all bacteria have the same exact genes

● Variations can be inherited → bacteria with antibiotic resistance traits (at random!) can

reproduce

● Organisms have more offspring that can survive under normal circumstances → bacteria

exponentially grow, favorably genes will be duplicated rapidly

● Variations that increase reproductive success will have a greater chance of being passed

on from generation to generation → bacteria with antibiotic resistance genes are more

likely to survive and reproduce

● A population will slowly change over time in response to the environment → this is why

antibiotic resistance is a problem, there are resistant species emerging, not just

individual colonies

So, really, where is

this coming from?

ABR Sources

A brief history...

Select Germs Showing Resistance Over Time...

Antibiotic

Year

Released

Drug Target

Resistant Germ Identified

Year

Identified

Penicillin

1941

Bacterial cells with

peptidoglycan cell

walls

Penicillin

-resistant

Staphylococcus aureus

Penicillin

-resistant

Streptococcus pneumoniae

1942

1967

Methicillin

1960

Inhibiting bacterial

cell

-wall synthesis,

similar to Penicillin

Methicillin

-resistant

Staphylococcus aureus

1960

Fluconazole

1988

Inhibits fungal

enzyme required for

fungal cell wall

synthesis.

Fluconazole

-resistant Candida

1990

Vocabulary Alert!

Peptidoglycan

What is it???

How do successful antibiotics work and what is the basis of

resistance to them?

They are overusedPeople are misusing them

The Problem with Current Antibiotics….

Don’t stop when you “feel better.”

Finish the entire course.

Listen to your doctor!

Overprescribed, sometimes even

asked for when not needed.

They rely on similar mechanisms of action

Targeting the peptidoglycan wall

(for example)

Antibiotic Misuse results in….

Our Plan

● Providing alternative selectable markers that do not rely on antibiotic resistance

genes

○ Alternative selectable markers are genes we can alter to produce obvious,

observable effects.

○ They allow us to determine which cells have been genetically altered and

which have not.

● How to test?

○ Give bacteria a plasmid of interest with our DNA of interest (via conjugation)

○ Cells that have taken up DNA of interest (with an antibiotic resistance marker

on it) will be able to survive when plated on the respective antibiotic while

those who have not taken up the DNA of interest will die when plated on the

antibiotic

○ This is important in bacterial transformations because we want to see if a cell

took up a plasmid of interest

Our Plan continued....

● We will be testing for carbon sources as positive selection markers

○ Sucrose, arabitol, and ribitol → see if we can transform cells and give

them the ability to metabolize these sugars → will be able to test for

transformants

03 Our Project Part - The Science Stuff!

Pseudomonas putida E. Coli

Bacteria we are using

Sugars we are working with...

Sucrose Arabitol Glucose Ribitol

Wet lab has

been busy...

31 2 3 4 5

7 8 9 10 11 12

14 15 16 17 18 19

21 22 23 24 25 26

28 29 30 31 1 2

130

Research

TestTestTest

Meeting

Lab Day

Lab Day Lab Day Lab Day

Lab Day

Our progress!

So far we have….

● Made plasmids with each of the four sugar pathways

● Made each plasmid with a backbone including antibiotic resistance

● Prepared plates with our sugars and put our bacteria on their plates

with corresponding inducers

METHODOLOGY

and

EQUIPMENT

Venus has a beautiful

name and it’s hot

Jupiter is a bright object

in the night sky

Saturn is composed of

hydrogen and helium

Pipettes, Centrifuges, Autoclaves, Plates

PCR

Gel Electrophoresis

PCR

Polymerase Chain Reaction

So how does PCR

work?

GEL

ELECTROPHORESIS

Gel Electrophoresis

continued….

The end result!

EQUIPMENT

Just the basics….

Pipettes

Plates

Centrifuge

Autoclave

Just to wrap it all

together again

Why this is important….

HOW YOU CAN GET

INVOLVED!

We’re holding an infographic/meme competition!

● Send us your best memes, artwork, or infographics related to iGEM,

synthetic biology, or antibiotic resistance, and we will credit you on our

iGEM wiki (if you want)!

● If your work is selected for publishing, you can put on your resume that

you assisted with a University of Florida iGEM team project, and put our

names and wiki URL as a reference!

● Top 3 winners get to contact us for advice related to UF, iGEM, science

research, and get a Zoom lab tour!

● Rules: unlimited submissions, anything related to this presentation, and

make sure it is funny or artistic (whatever you are going for)

Submission Instructions

● Email: ufigemteam@gmail.com

● Subject line: RISE project

● Send in highest resolution possible! We accept PNG, TIFF, JPG, PDF, etc.

Examples

CONCLUSIONS

Antibiotic Resistance is coming! Do your part to

complete your dose and listen to your doctor!!!

● Science is fun! But complicated!

● Keep iGEM and UF on your radar in the future!

your submissions!

@uf.igem